phi29 DNA Polymerase |NEB酶试剂 New England Biolabs

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产品信息

phi29 DNA Polymerase is the replicative polymerase from the Bacillus subtilis phage phi29 (Φ29) (1). This polymerase has exceptional strand displacement and processive synthesis properties (2). The polymerase has an inherent 3´→5′ proofreading exonuclease activity (3).

重点

 

产品来源

An E. coli strain that carries the phi29 DNA Polymerase gene from bacteriophage phi29

产品类别:
Isothermal Amplification & Strand Displacement Products

应用:
Whole Genome Amplification & Multiple Displacement Amplification,
Isothermal Amplification

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • M0269S     -20    
        phi29 DNA Polymerase M0269SVIAL -20 1 x 0.025 ml 10,000 units/ml
        phi29 DNA Polymerase Reaction Buffer B0269SVIAL -20 1 x 1.5 ml 10 X
        Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml
    • M0269L     -20    
        phi29 DNA Polymerase M0269LVIAL -20 1 x 0.125 ml 10,000 units/ml
        phi29 DNA Polymerase Reaction Buffer B0269SVIAL -20 1 x 1.5 ml 10 X
        Recombinant Albumin, Molecular Biology Grade B9200SVIAL -20 1 x 0.6 ml 20 mg/ml

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme that will incorporate 0.5 pmol of dNTP into acid insoluble material in 10 minutes at 30°C.

    反应条件

    1X phi29 DNA Polymerase Reaction Buffer
    Supplement with Recombinant Albumin, Molecular Biology Grade
    Incubate at 30°C

    1X phi29 DNA Polymerase Reaction Buffer
    50 mM Tris-HCl
    10 mM MgCl2
    10 mM (NH4)2SO4
    4 mM DTT
    (pH 7.5 @ 25°C)

    贮存溶液

    10 mM Tris-HCl
    100 mM KCl
    1 mM DTT
    0.1 mM EDTA
    50% Glycerol
    0.5% Tween® 20
    0.5% IGEPAL® CA-630
    pH 7.4 @ 25°C

    热失活

    65°C for 10 minutes

    分子量

    理论上的: 67000 daltons

    5′ – 3′ 核酸外切酶

    No

    3′ – 5′ 核酸外切酶

    Yes

    链置换

    ++++

    单位活性检测条件

    1X phi29 DNA Polymerase Reaction Buffer, 0.1 mg/ml Recombinant Albumin, 0.01 mg/ml HindIII-digested λ DNA, 0.2 µM dTTP including [3H]-dTTP, 0.2 mM dGTP, 0.2 mM dATP and 0.2 mM dCTP.

  • 优势和特性

    应用特性

    • Replication requiring a high degree of strand displacement and/or processive synthesis
    • High fidelity replication at moderate temperatures

  • 相关产品

    相关产品

    • dNTP 混合液
    • dNTP 套装

    单独销售的组分

    • 重组白蛋白, 分子生物学级（不含动物成分）

  • 注意事项

    1. The presence of active reducing reagent in the reaction buffer is critical for this enzyme. While the reaction buffer supplied with the enzyme contains DTT, older buffer stocks or stocks that have been repeatedly frozen and thawed should be supplemented with 4 mM DTT to obtain maximal activity.

  • 参考文献

    1. Blanco, L. and Salas, M. (1984). Proc. Natl. Acad. Sci. USA. 81, 5325-5329.
    2. Blanco. L., et al. (1989). J. Biol. Chem.. 264, 8935-8940.
    3. Garmendia, C., et al. (1992). J. Biol. Chem.. 267, 2594-2599.

操作说明、说明书 & 用法

  • 使用指南

    • Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

工具 & 资源

  • 选择指南

    • DNA Polymerase Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. Can phi29 DNA Polymerase be used in other NEBuffers?
    2. Can phi29 DNA Polymerase be used to blunt DNA?
    3. Can phi29 DNA Polymerase be used to fill in 3′ overhangs?
    4. Can phi29 DNA Polymerase be used to remove 5′ overhangs?
    5. Can phi29 DNA Polymerase be heat inactivated?
    6. At what rate does phi29 DNA Polymerase add nucleotides to a primed single-stranded template?
    7. Will phi29 DNA Polymerase work in rCutSmart buffer?
    8. Are NEB DNA Polymerases supplied with dNTPs?

  • 实验技巧

    DTT in the reaction buffer can oxidize quickly with >-20°C storage temperatures, extended storage, or multiple freeze/thaws, which can reduce amplification efficiency of phi29 DNA Polymerase.  Older buffer stocks or stocks that have been repeatedly frozen and thawed should be supplemented with 4 mM DTT to obtain maximal activity.